Leucophor 1111X

Endpoint summary

Description of key information

not skin irritating
not eye irritating

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

From August 24th to September 06th, 2005

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Study conducted according to internationally accepted testing guidelines and performed in compliance with Good Laboratory Practice. Justification for read across approach is given in the endpoint summary and in the read across justification report attached to the Section 13 of this dossier.

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

adopted April 24, 2002

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Version / remarks:

April 29, 2004

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland GmbH Stolzenseeweg 32-36 D-88353 Kisslegg / Germany.
- Age at study initiation: 11-12 weeks (male); 10-11 weeks (females).
- Weight at study initiation: 1687 - 2454 g
- Housing: individually in stainless steel cages equipped with feed hoppers and drinking water bowls. Wood blocks and haysticks 4642 (batch no. 35/05, Provimi Kliba AG) were provided for gnawing.
- Diet: pelleted standard Provimi Kliba 3418 rabbit maintenance diet ad libitum (batch no. 34/05) provided by Provimi Kliba AG, CH-4303 Kaiseraugst.
- Water: community tap water from Füllinsdorf, ad libitum.
- Acclimation period: under laboratory conditions after health examination. Only animals without any visual signs of illness were used for the study.

ENVIRONMENTAL CONDITIONS
- Temperature: ranges for room temperature 17-23 °C.
- Humidity: 30-70 %
- Air changes: approximately 10-15 air changes per hour.
- Photoperiod: animals were provided with an automatically controlled light cycle of 12 hours light and 12 hours dark.
- Other: music was played during the daytime light period.
Room temperature and humidity were monitored continuously and values outside of these ranges may have occasionally occurred, usually following room cleaning. These transient variations are considered not to have any influence on the study and, therefore, these data are not reported but are retained at testing laboratory.

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

water

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST ITEM PREPARATION
0.5 g (per animal) of the test item was weighed as delivered by the Sponsor and then moistened with approximately 0.5 ml of purified water before application.
The pH of the test item was measured before the study initiation date. A formulation of a 1 % (w/w) solution was prepared. The pH was found to be 8.07.

Duration of treatment / exposure:

4 hours

Observation period:

7 days

Number of animals:

3 (animals of both sexes were used)

Details on study design:

As it was suspected that the test item might produce irritancy, a single animal (one female) was treated first. As neither a corrosive effect nor a severe irritant effect was observed after the 4-hour exposure, the test was completed using the two remaining animals for an exposure period of four hours.

TEST SITE
- Area of exposure: four days before treatment, the left flank was clipped with an electric clipper, exposing an area of approximately 100 cm2 (10 cm x 10 cm). The skin of the animals was examined one day before treatment, and regrown fur of all animals was clipped again. Animals with overt signs of skin injury or marked irritation which may have interfered with the interpretation of the results were not used in the test.
- Type of wrap if used: on the day of treatment, 0.5 g of test item was placed on a surgical gauze patch (ca. 4 cm x 4 cm). The patch was covered with a semi-occlusive dressing. The dressing was wrapped around the abdomen and anchored with tape.

REMOVAL OF TEST SUBSTANCE
- Washing: the dressing was removed and the skin was flushed with lukewarm tap water to clean the application site. The test item could not be removed completely, so that remnants were still present at the 1-hour reading in all animals. However, any reactions (erythema) were clearly visible at that time.
- Time after start of exposure: 4 hours.

OBSERVATIONS
- Viability/Mortality: daily from acclimatization of the animals to the termination of test.
- Clinical signs: daily from acclimatization of the animals to the termination of test.
- Body weights: at start of acclimatization, on the day of application and at termination of observation.

NECROPSY
No necropsy was performed on the animals sacrificed at termination of observation.
All rabbits were sacrificed by an intravenous injection of Vetanarcol into the ear vein at a dose of at least 1 ml/kg body weight (equivalent to 162 mg sodium pentobarbitone/kg body weight) and discarded.

SCORING SYSTEM
The skin reaction was assessed according to the numerical scoring system listed in the Commission Directive 2004/73/EC, April 29, 2004, approximately 1, 24, 48 and 72 hours, as well as 7 days after the removal of the dressing, gauze patch and test item.
If evident, corrosive or staining properties of the test item were described and recorded.

Grading of Skin Reactions
ERYTHEMA AND ESCHAR FORMATION
No erythema: 0
Very slight erythema: 1
Well-defined erythema: 2
Moderate to severe erythema: 3
Severe erythema (beet redness) or eschar formation (injuries in depth preventing erythema) reading: 4

OEDEMA FORMATION
No oedema: 0
Very slight oedema (barely perceptible): 1
Slight oedema (edges of area well-defined by definite raising): 2
Moderate oedema (edges raised approximately 1 mm): 3
Severe oedema (raised more than 1 mm and extending beyond the area of exposure): 4

Irritation parameter:

erythema score

Basis:

animal: 3/3

Time point:

other: 24, 28 and 72 hrs

Score:

< 2.3

Irritation parameter:

edema score

Basis:

animal: 3/3

Time point:

other: 24, 28 and 72 hrs

Score:

< 2.3

Irritant / corrosive response data:

The mean score was calculated across 3 scoring times (24, 48 and 72 hours after patch removal) for each animal for erythema/eschar grades and for oedema grades, separately. The mean erythema/eschar score of the three animals was 0.67, 1.00 and 1.33, respectively and the mean oedema score was 0.00 for all three animals, respectively.
Well-defined to very slight erythema was noted in all animals from the 1- to the 48-hour reading and very slight erythema persisted in two animals up to the 72-hour observation.
Very slight swelling was observed in two animals at the 1-hour reading.
Scaling was present in one animal at the 72-hour reading.
No abnormal findings were observed on the treated skin of any animal 7 days after treatment, the end of the observation time.

COLORATION
No staining produced by the test item of the treated skin was observed.

CORROSION
Neither alterations of the treated skin were observed nor were corrosive effects evident on the skin.

Other effects:

No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred.
The body weights of all rabbits were considered to be within the normal range of variability.

SKIN IRRITATION SCORES

Animal N.	Sex	Reaction	Evaluation interval					Mean 24, 48 and 72 hrs
			1 hr	24 hrs	48 hrs	72 hrs	7 days
19	M	Erythema	2	1	1	0	0	0.67
20	F	Erythema	2	1	1	1	0	1.00
21	F	Erythema	2	2	1	1	0	1.33
19	M	Oedema	1	0	0	0	0	0.00
20	F	Oedema	0	0	0	0	0	0.00
21	F	Oedema	1	0	0	0	0	0.00
19	M	Scaling	0	0	0	0	0
20	F	Scaling	0	0	0	0	0
21	F	Scaling	0	0	0	X	0

x = scaling present

INDIVIDUAL FINDINGS

	ANIMAL N. 19, MALE	ANIMAL N. 20, FEMALE	ANIMAL N. 21, FEMALE
After 1 hour	Erythema: well-defined erythema Oedema: very slight swelling Flaking: no abnormal findings noted Staining: no abnormal findings noted Other: remnants of the test item present	Erythema: well-defined erythema Oedema: no abnormal findings noted Flaking: no abnormal findings noted Staining: no abnormal findings noted Other: remnants of the test item present	Erythema: well-defined erythema Oedema: very slight swelling Flaking: no abnormal findings noted Staining: no abnormal findings noted Other: remnants of the test item present
After 24 hours	Erythema: very slight erythema Oedema: no abnormal findings noted Flaking: no abnormal findings noted Staining: no abnormal findings noted	Erythema: very slight erythema Oedema: no abnormal findings noted Flaking: no abnormal findings noted Staining: no abnormal findings noted	Erythema: well-defined erythema Oedema: no abnormal findings noted Flaking: no abnormal findings noted Staining: no abnormal findings noted
After 48 hours	Erythema: very slight erythema Oedema: no abnormal findings noted Flaking: no abnormal findings noted Staining: no abnormal findings noted	Erythema: very slight erythema Oedema: no abnormal findings noted Flaking: no abnormal findings noted Staining: no abnormal findings noted	Erythema: very slight erythema Oedema: no abnormal findings noted Flaking: no abnormal findings noted Staining: no abnormal findings noted
After 72 hours	Erythema: no abnormal findings noted Oedema: no abnormal findings noted Flaking: no abnormal findings noted Staining: no abnormal findings noted	Erythema: very slight erythema Oedema: no abnormal findings noted Flaking: no abnormal findings noted Staining: no abnormal findings noted	Erythema: very slight erythema Oedema: no abnormal findings noted Flaking: scaling present Staining: no abnormal findings noted
After 7 days	Erythema: no abnormal findings noted Oedema: no abnormal findings noted Flaking: no abnormal findings noted Staining: no abnormal findings noted	Erythema: no abnormal findings noted Oedema: no abnormal findings noted Flaking: no abnormal findings noted Staining: no abnormal findings noted	Erythema: no abnormal findings noted Oedema: no abnormal findings noted Flaking: no abnormal findings noted Staining: no abnormal findings noted

Interpretation of results:

not classified

Remarks:

Migrated information according to the CLP Regulation (EC 1272/2008) Criteria used for interpretation of results: EU

Conclusions:

not irritating

Executive summary:

The primary skin irritation potential of the substance was investigated according to OECD test guideline no. 404. The test item was applied by topical semi-occlusive application of 0.5 g to the intact left flank of each of three young adult New Zealand White rabbits. The duration of treatment was four hours. The scoring of skin reactions was performed 1, 24, 48 and 72 hours, as well as 7 days after removal of the dressing.

The mean score was calculated across 3 scoring times (24, 48 and 72 hours after patch removal) for each animal for erythema/eschar grades and for oedema grades, separately. The mean erythema/eschar score of the three animals was 0.67, 1.00 and 1.33, respectively and the mean oedema score was 0.00 for all three animals, respectively.

The application of test item to the skin resulted in mild signs of irritation. These included erythema, oedema and scaling. These effects were reversible and were no longer evident 7 days after treatment, the end of the observation period for all animals. The test item caused no staining of the treated skin, but could not be removed completely by washing with lukewarm water at the removal of the dressing, so that remnants were still present at the 1-hour reading in all animals. However, any reactions (erythema) were clearly visible at that time. No corrosive effects were noted on the treated skin of any animal at any of the measuring intervals and no clinical signs were observed. Thus, the test item did not induce significant or irreversible damage to the skin.

Conclusion

Mean values from gradings at 24, 48 and 72 hours after patch removal were lower than 2.3 in all animals for both erythema/eschar and oedema reactions.

In conclusion, the test item does not meet the criteria ti be classified as irritating, according to the CLP (EC 1272/2008) Regulation.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

From September 07th to 16th, 2005

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Study conducted according to internationally accepted testing guidelines and performed in compliance with Good Laboratory Practice. Justification for read across approach is given in the endpoint summary and in the read across justification report attached to the Section 13 of this dossier.

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

adopted April 24, 2002

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Version / remarks:

April 29, 2004

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland GmbH Stolzenseeweg 32-36 D-88353 Kisslegg / Germany.
- Age at study initiation: 12-14 weeks.
- Weight at study initiation: 1687 - 2454 g
- Housing: individually in stainless steel cages equipped with feed hoppers and drinking water bowls. Wood blocks and haysticks 4642 (batch no. 35/05, Provimi Kliba AG) were provided for gnawing.
- Diet: pelleted standard Provimi Kliba 3418 rabbit maintenance diet ad libitum (batch no. 34/05) provided by Provimi Kliba AG, CH-4303 Kaiseraugst.
- Water: community tap water from Füllinsdorf, ad libitum.
- Acclimation period: under laboratory conditions after health examination. Only animals without any visual signs of illness were used for the study.

ENVIRONMENTAL CONDITIONS
- Temperature: ranges for room temperature 17-23 °C.
- Humidity: 30-70 %
- Air changes: approximately 10-15 air changes per hour.
- Photoperiod: animals were provided with an automatically controlled light cycle of 12 hours light and 12 hours dark.
- Other: music was played during the daytime light period.
Room temperature and humidity were monitored continuously and values outside of these ranges may have occasionally occurred, usually following room cleaning. These transient variations are considered not to have any influence on the study and, therefore, these data are not reported but are retained at testing laboratory.

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.1 g (per animal) was weighed and applied undiluted as it was delivered by the Sponsor.
- pH: the pH of 1 % (w/w) solution of the test item was measured for previous study and was found to be 8.07.

Number of animals or in vitro replicates:

3 (animals of both sexes were used)

Details on study design:

The eyes of the animals were examined one day prior to test item administration.
Animals with overt signs of ocular injury or irritation which may have interfered with the interpretation of the results were not used in the test.

APPLICATION OF TEST MATERIAL
- Application: on the day of treatment, 0.1 g of the test item was placed in the conjunctival sac of the left eye of each animal after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second to prevent loss of test item.
- Control: the right eye remained untreated and served as the reference control.
As it was suspected that the test item might produce irritancy, a single animal (one female) was treated first. As neither a corrosive effect nor a severe irritant effect was observed after the 1- and 24-hour examinations, the test was completed using the two remaining animals.

REMOVAL OF TEST SUBSTANCE
- Washing: the treated eyes were not rinsed after instillation.

OBSERVATIONS
- Viability/Mortality: daily from acclimatization of the animals to the termination of test.
- Clinical signs: daily from acclimatization of the animals to the termination of test.
- Body weights: at start of acclimatization, on the day of application and at termination of observation.

NECROPSY
No necropsy was performed on the animals sacrificed at termination of observation.
All rabbits were sacrificed by an intravenous injection of Vetanarcol into the ear vein at a dose of at least 1 ml/kg body weight (equivalent to 162 mg sodium pentobarbitone/kg body weight) and discarded.

TOOL USED TO ASSESS SCORE
Eye examinations were made with a Varta Cliptrix diagnostic-lamp (Roth AG, CH-4153 Reinach/Switzerland).

SCORING SYSTEM
The ocular reaction (i.e. corneal opacity, iridic effects, conjunctivae and chemosis) was assessed according to the numerical scoring system listed in the Commission Directive 2004/73/EC, April 29, 2004, at approximately 1, 24, 48 and 72 hours after instillation.
Additionally, ocular discharge, reddening of the sclerae and staining of conjunctivae, sclerae and cornea by the test item was assessed.

Grading of Ocular Lesions
CORNEA
Opacity: degree of density (area most dense taken for reading)
No ulceration or opacity: 0
Scattered or diffuse areas of opacity (other than slight dulling of normal lustre), details of iris clearly visible: 1
Easily discernible translucent area, details of iris slightly obscured: 2
Nacreous area, no details of iris visible, size of pupil barely discernible: 3
Opaque comea, iris not discernible through the opacity: 4
Area of cornea involved
Zero: 0
One quarter (or less) but not zero: 1
Greater than one quarter, but less than half: 2
Greater than half, but less than three quarters: 3
Greater than three quarters, up to whole area: 4

IRIS
Normal: O
Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia, or injection, any of these or combination of any thereof, iris still reacting to light (sluggish reaction is positive): 1
No reaction to light, haemorrhage, gross destruction (any or all of these): 2

CONJUNCTIVAE
Redness (refers to most severe reading of palpebral and bulbar conjunctivae when compared with control eye)
Blood vessels normal: 0
Some blood vessels definitely hyperaemic (injected): 1
Diffuse, crimson colour, individual vessels not easily discernible: 2
Diffuse beefy red: 3
Chemosis: lids and/or nictitating membranes
No swelling: 0
Any swelling above normal (including nictitating membranes): 1
Obvious swelling with partial eversion of lids: 2
Swelling with lids about half-closed: 3
Swelling with lids more than half-closed: 4

ADDITIONALLY, OCULAR DISCHARGE, REDDENING OF THE SCLERAE AND STAINING OF CONJUNCTIVAE, SCLERAE AND CORNEA BY THE TEST ITEM WAS ASSESSED ACCORDING TO THE FOLLOWING SCHEME:
OCULAR DISCHARGE
No discharge: 0
Slight: any amount different to normal (does not include small amount observed in inner canthus of normal animal): 1
Moderate: discharge with moistening of the lids and hair just adjacent to the lids: 2
Marked: discharge with moistening of the lids and hairs, and a considerable area around the eye (running): 3

SCLERAE
Redness
Normal: blood vessels nomial: 0
Slight reddening: some blood vessels definitely hyperemic (injected): 1
Moderate reddening: diffuse, crimson colour, individual vessels not easily disoernible: 2
Marked reddening: diffuse beefy red: 3

STAINING OF CONJUNCTIVAE, SCLERAE AND CORNEA BY THE TEST ITEM
Not observed: 0
Slight staining: 1
Marked staining: 2

Irritation parameter:

cornea opacity score

Basis:

animal: 3/3

Time point:

other: 24, 28 and 72 hrs

Score:

< 1

Irritation parameter:

iris score

Basis:

animal: 3/3

Time point:

other: 24, 28 and 72 hrs

Score:

< 1

Irritation parameter:

conjunctivae score

Basis:

animal: 3/3

Time point:

other: 24, 28 and 72 hrs

Score:

< 2

Irritation parameter:

chemosis score

Basis:

animal: 3/3

Time point:

other: 24, 28 and 72 hrs

Score:

< 2

Irritant / corrosive response data:

The mean score was calculated across 3 scoring times (24, 48 and 72 hours after instillation) for each animal for corneal opacity, iris, redness and chemosis of the
conjunctivae, separately. The individual mean scores for corneal opacity and iris were 0.00 for all three animals. The individual mean scores for the conjunctivae were 0.00, 0.33 and 0.33 for reddening and 0.00 for chemosis for all three animals, respectively.
No abnormal Endings were observed in the cornea or iris of any animal at any of the measurement intervals.
Slight to moderate reddening of the conjunctivae was noted in all animals at the 1-hour reading and slight reddening persisted in two animals up to 24 hours after test item instillation.
Slight swelling of the conjunctivae (chemosis) to obvious swelling with partial eversion of the lids was observed in two animals one hour after treatment.
Slight to moderate reddening of the sclerae was present in all animals at the 1-hour observation and slight reddening persisted in one animal up to the 24-hour examination.
Slight ocular discharge was observed in one animal one hour after treatment.
No abnormal findings were observed in the treated eye of any animal 48 hours after treatment.

COLORATION
No staining of the treated eyes produced by the test item was observed.

CORROSION
No corrosion of the cornea was observed at any of the reading times.

Other effects:

No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred.

EYE IRRITATION SCORES

Animal	Sex	1 hr	24 hrs	48 hrs	72 hrs	Mean 24, 48, 72 hrs
Cornea opacity
22	M	0	0	0	0	0
23	F	0	0	0	0	0
24	F	0	0	0	0	0
Iritis
22	M	0	0	0	0	0
23	F	0	0	0	0	0
24	F	0	0	0	0	0
Conjunctival Redness
22	M	1	0	0	0	0
23	F	2	1	0	0	0.33
24	F	2	1	0	0	0.33
Conjunctival Chemosis
22	M	0	0	0	0	0
23	F	2	0	0	0	0
24	F	1	0	0	0	0
Sclera
22	M	1	0	0	0
23	F	2	1	0	0
24	F	2	0	0	0

INDIVIDUAL FINDINGS

	ANIMAL N. 22, MALE	ANIMAL N. 23, FEMALE	ANIMAL N. 24, FEMALE
After 1 hour	Cornea: no abnormal findings noted Iris: no abnormal findings noted Conjunctivae: slightly reddened Discharge: no abnormal findings noted Slerae: slightly reddened Test item: no remnants evident	Cornea: no abnormal findings noted Iris: no abnormal findings noted Conjunctivae: moderately reddened, obvious swelling with partial eversion of the lids Discharge: no abnormal findings noted Slerae: moderately reddened Test item: no remnants evident	Cornea: no abnormal findings noted Iris: no abnormal findings noted Conjunctivae: moderately reddened, slight swelling Discharge: slight discarge Slerae: moderately reddened Test item: no remnants evident
After 24 hours	Cornea: no abnormal findings noted Iris: no abnormal findings noted Conjunctivae: no abnormal findings noted Discharge: no abnormal findings noted Slerae: no abnormal findings noted Test item: no remnants evident	Cornea: no abnormal findings noted Iris: no abnormal findings noted Conjunctivae: slightly reddened Discharge: no abnormal findings noted Slerae: slightly reddened Test item: no remnants evident	Cornea: no abnormal findings noted Iris: no abnormal findings noted Conjunctivae: slightly reddened Discharge: no abnormal findings noted Slerae: no abnormal findings noted Test item: no remnants evident
After 48 hours	Cornea: no abnormal findings noted Iris: no abnormal findings noted Conjunctivae: no abnormal findings noted Discharge: no abnormal findings noted Slerae: no abnormal findings noted Test item: no remnants evident	Cornea: no abnormal findings noted Iris: no abnormal findings noted Conjunctivae: no abnormal findings noted Discharge: no abnormal findings noted Slerae: no abnormal findings noted Test item: no remnants evident	Cornea: no abnormal findings noted Iris: no abnormal findings noted Conjunctivae: no abnormal findings noted Discharge: no abnormal findings noted Slerae: no abnormal findings noted Test item: no remnants evident
After 72 hours	Cornea: no abnormal findings noted Iris: no abnormal findings noted Conjunctivae: no abnormal findings noted Discharge: no abnormal findings noted Slerae: no abnormal findings noted Test item: no remnants evident	Cornea: no abnormal findings noted Iris: no abnormal findings noted Conjunctivae: no abnormal findings noted Discharge: no abnormal findings noted Slerae: no abnormal findings noted Test item: no remnants evident	Cornea: no abnormal findings noted Iris: no abnormal findings noted Conjunctivae: no abnormal findings noted Discharge: no abnormal findings noted Slerae: no abnormal findings noted Test item: no remnants evident

Interpretation of results:

not classified

Remarks:

Migrated information according to the CLP Regulation (EC 1272/2008) Criteria used for interpretation of results: EU

Conclusions:

not irritating

Executive summary:

The primary eye irritation potential of test item was investigated according to OECD test guideline no. 405. The test item was applied by instillation of 0.1 g into the left eye of each of three young adult New Zealand White rabbits. Scoring of irritation effects was performed approximately 1, 24, 48 and 72 hours after test item instillation. The mean score was calculated across 3 scoring times (24, 48 and 72 hours after instillation) for each animal for corneal opacity, iris, redness and chemosis of the conjunctivae, separately. The individual mean scores for corneal opacity and iris were 0.00 for all three animals. The individual mean scores for the conjunctivae were 0.00, 0.33 and 0.33 for reddening and 0.00 for chemosis for all three animals, respectively. The instillation of the substance into the eye resulted in mild, early-onset and transient ocular changes, such as reddening of the conjunctivae and sclerae, discharge and chemosis. These effects were reversible and were no longer evident 48 hours after treatment. No abnormal Endings were observed in the cornea or iris of any animal at any of the examinations. No corrosion was observed at any of the measuring intervals. No staining of the treated eyes by the test item was observed and no clinical signs were observed. Thus, the test item did not induce significant or irreversible damage to the rabbit eye.

Conclusion

The mean values from gradings at 24, 48 and 72 hours were lower than 1 for corneal opacity, lower than 1 for irtitis, lower than 2 for both conjunctival redness and oedema, in all animals.

Therefore the substance does not meet the criteria to be classified as eye irritating, according to the CLP (EC 1272/2008) Regulation.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

There are no experimental data available on the Leucophor 1111X, thus a read across approach with the structural analogous substances has been proposed.

All the substances used in a read across approach are Stilbene derivatives Fluorescent Whitening Agents salts. They display similar structural and physicochemical properties; all of them exhibit high degree of dissociation in water and very low octanol/water partition coefficients because to a higher affinity with water phase than the octanol one. The water solubility is due to the sulphonated groups in the molecules.

The differences occurring in the structure formulas (i.e. substituents) are expected to not significantly impact the toxicological characterisation.

Further details about the justification for read across approach are given in the report attached to the Section 13 of this dossier.

SKIN IRITATION

The primary skin irritation potential of the analogous substance 01 was investigated according to OECD test guideline no. 404. The test item was applied by topical semi-occlusive application of 0.5 g to the intact left flank of each of three young adult New Zealand White rabbits. The application of test item to the skin resulted in mild signs of irritation: these included erythema, oedema and scaling. These effects were reversible and were no longer evident 7 days after treatment, the end of the observation period for all animals. The test item caused no staining of the treated skin, but could not be removed completely by washing with lukewarm water at the removal of the dressing, so that remnants were still present at the 1-hour reading in all animals. However, any reactions (erythema) were clearly visible at that time. No corrosive effects were noted on the treated skin of any animal at any of the measuring intervals and no clinical signs were observed. Thus, the test item did not induce significant or irreversible damage to the skin. The mean values from gradings at 24, 48 and 72 hours after patch removal were lower than 2.3 in all animals for both erythema/eschar and oedema reactions, thus the test item does not meet the criteria to be classified as irritating, according to the CLP (EC 1272/2008) Regulation (Bertl, 2005).

This result is confirmed by the outcomes from the available studies on the other analogous (analogous substance 02 and analogous substance 05).

EYE IRRITATION

The primary eye irritation potential of the analogous substance 01 was investigated according to OECD test guideline no. 405. The test item was applied by instillation of 0.1 g into the left eye of each of three young adult New Zealand White rabbits. The instillation of the substance into the eye resulted in mild, early-onset and transient ocular changes, such as reddening of the conjunctivae and sclerae, discharge and chemosis. These effects were reversible and were no longer evident 48 hours after treatment. No abnormal Endings were observed in the cornea or iris of any animal at any of the examinations. No corrosion was observed at any of the measuring intervals. No staining of the treated eyes by the test item was observed and no clinical signs were observed. The mean values from gradings at 24, 48 and 72 hours were lower than 1 for corneal opacity, lower than 1 for irtitis, lower than 2 for both conjunctival redness and oedema, in all animals, thus the substance can be considered as non irritating to eye (Bertl, 2005).

This result is confirmed by the outcomes from the available studies on the other analogous (analogous substance 02 and analogous substance 05).

Justification for selection of skin irritation / corrosion endpoint:
Study conducted according to internationally accepted testing guidelines and performed in compliance with Good Laboratory Practice.

Justification for selection of eye irritation endpoint:
Study conducted according to internationally accepted testing guidelines and performed in compliance with Good Laboratory Practice.

Justification for classification or non-classification

According to the CLP Regulation (EC 1272/2008), 3.2 Skin corrosion/irritation section, skin Irritation means the production of reversible damage to the skin following the application of a test substance for up to 4 hours.

In the key study, the mean values from gradings at 24, 48 and 72 hours after patch removal were lower than 2.3 in at least 2 of 3 animals for both erythema/eschar and oedema reactions.

According to the CLP Regulation (EC 1272/2008) substances that have the potential to induce reversible eye irritation are classified in Category 2 (irritating to eyes) if, when applied to the eye of an animal, a substance produces at least in 2 of 3 tested animals, a positive response of corneal opacity ≥ 1 and/or iritis ≥ 1, and/or conjunctival redness ≥ 2 and/or conjunctival oedema (chemosis) ≥ 2, calculated as the mean scores following grading at 24, 48 and 72 hours after installation of the test material, and which fully reverses within an observation period of 21 days.

The mean values from gradings at 24, 48 and 72 hours were lower than 1 for corneal opacity, lower than 1 for iritis, lower than 2 for both conjunctival redness and oedema, in all animals in the key study.

In conclusion, the substance is not classified for the eye/skin irritation, according to the CLP Regulation (EC 1272/2008).